Controlled release peptide formulations

ABSTRACT

The present invention relates to compositions forming a low viscosity mixture of:a)20-80 wt. % of at least one diacyl glycerol and/or a tocopherol;b)20-80 wt. % of at least one phosphatidyl choline (PC);c)5-20 wt. % of at least one biocompatible, organic mono-alcoholicsolvent;d)up to 20 wt. % polar solvente)at least one peptide active agent;f)optionally at least one antioxidant;wherein the ratio of components a:b is in the range 40:60 to 54:46;wherein the pre-formulation forms, or is capable of forming, at least one liquid crystalline phase structure upon contact with excess aqueous fluid.The invention further relates to methods of treatment comprising administration of such compositions, and to pre-filled administration devices and kits containing the formulations.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser.No. 14/117,994, filed on Feb. 20, 2014, which is a § 371 application ofInternational Application No. PCT/EP2012/059917, filed May 25, 2012,which claims priority to U.S. Application No. 61/489,886, filed on May25, 2011, each of which is incorporated herein by reference.

SEQUENCE LISTING

The Sequence Listing submitted herewith is an ASCII text file(2021-06-17_Sequence_Listing.text, created on Jun. 16, 2021, 559 bytes)via EFS-Web is hereby incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to formulation precursors(pre-formulations) for the in situ generation of compositions for thecontrolled release of peptide active agents, and methods of treatmentwith such formulations. In particular, the invention relates tohigh-loading pre-formulations of amphiphilic components and at least onepeptide active agent for parenteral application, which undergo phasetransition upon exposure to aqueous fluids, such as body fluids, therebyforming a controlled release composition.

BACKGROUND TO THE INVENTION

Many bioactive agents including pharmaceuticals, nutrients, vitamins andso forth have a “functional window”. That is to say that there is arange of concentrations over which these agents can be observed toprovide some biological effect. Where the concentration in theappropriate part of the body (e.g. locally or as demonstrated by serumconcentration) falls below a certain level, no beneficial effect can beattributed to the agent. Similarly, there is generally an upperconcentration level above which no further benefit is derived byincreasing the concentration. In some cases increasing the concentrationabove a particular level results in undesirable or even dangerouseffects.

Some bioactive agents have a long biological half-life and/or a widefunctional window and thus may be administered occasionally, maintaininga functional biological concentration over a substantial period of time(e.g. 6 hours to several days). In other cases the rate of clearance ishigh and/or the functional window is narrow and thus to maintain abiological concentration within this window regular (or even continuous)doses of a small amount are required. This can be particularly difficultwhere non-oral routes of administration (e.g. parenteral administration)are desirable or necessary, since self-administration may be difficultand thus cause inconvenience and/or poor compliance. In such cases itwould be advantageous for a single administration to provide activeagent at a therapeutic level over the whole period during which activityis needed.

There is an enormous potential in the use of peptides (includingproteins) for treating various disease states, as well as in prophylaxisand in improving general health and well-being of subjects. However, theperformance of administered peptide agents is generally limited due topoor bioavailability, which in turn is caused by the rapid degradationof peptides and proteins in biological fluids. This increases the dosewhich must be administered and in many cases restricts the effectiveroutes of administration. These effects are further exaggerated by theoften limited permeability of peptides and proteins across biologicalmembranes.

Peptides and proteins that are administered to the mammalian body (e.g.orally, intramuscularly etc.) are subject to degradation by variousproteolytic enzymes and systems present throughout the body. Well knownsites of peptidase activity include the stomach (e.g. pepsin), and theintestinal tract (e.g. trypsin, chymotrypsin, and others) but otherpeptidases (e.g. aminopeptidases, carboxypeptidases, etc.) are foundthroughout the body. Upon oral administration, gastric and intestinaldegradation reduces the amount of peptide or protein which potentiallycould be absorbed through the intestinal surface lining and therebydecreases their bioavailability. Similarly, free peptides and proteinsin the mammalian blood stream are also subject to enzymatic degradation(e.g. by plasma proteases etc.).

Some patients undergoing treatment will typically require a therapeuticdose to be maintained for a considerable period and/or ongoing treatmentfor many months or years. Thus a depot system allowing loading andcontrolled release of a larger dose over a longer period would offer aconsiderable advantage over conventional delivery systems.

Peptides may be delivered by systems such as the Alkermes Medisorb®delivery system consisting of microspheres of biodegradable polymers.Such polymer microsphere formulations must generally be administered bymeans of a sizable needle, typically of 20-gauge or wider. This isnecessary as a result of the nature of the polymeric dosing systemsused, which are typically polymer suspensions.

Evidently, it would be an advantage to provide a system of lowviscosity, such as a homogeneous solution, dispersion of fine particles,or L₂ phase, which could be administered easily through a narrow needle,thus decreasing the discomfort of the patient during the procedure. Thisease of administration is particularly significant where patients willbe on a self-administration regime and may already be self-administeringseveral times each day. Providing a sustained formulation with aduration of a few days, but which is sufficiently complex to administerthat it requires treatment by a healthcare professional will not be anadvantage to all patients over twice-daily or daily self-administration,and is likely to be more costly. Providing a formulation which givessufficiently long duration to justify a visit to a health professionalfor administration and/or a preparation which can be self-administered,and reducing preparation time of health-care professionals or patientsprior to the actual administration are all important issues.

The poly-lactate, poly-glycolate and poly-lactate-co-glycolate polymerstypically used for degrading slow-release formulations are also thecause of some irritation in at least some patients. In particular, thesepolymers typically contain a certain proportion of acetic acid impurity,which will irritate the injection site on administration. When thepolymer then breaks down, lactic acid and glycolic acid are thedegradation products so that further irritation is caused. As a resultof the combined effects of wide-needle administration and irritantcontents, discomfort at the site of administration and the formation ofconnective scar tissue are greater than desirable.

From a drug delivery point of view, polymer depot compositions generallyhave the disadvantage of accepting only relatively low drug loads andhaving a “burst/lag” release profile. The nature of the polymericmatrix, especially when applied as a solution or pre-polymer, causes aninitial burst of drug release when the composition is firstadministered. This is followed by a period of low release, while thedegradation of the matrix begins, followed finally by an increase in therelease rate to the desired sustained profile. This burst/lag releaseprofile can cause the in vivo concentration of active agent to burstabove the functional window immediately following administration, andthen drop back through the bottom of the functional window during thelag period before reaching a sustained functional concentration for aperiod of time. Evidently, from a functional and toxicological point ofview this burst/lag release profile is undesirable and could bedangerous. It may also limit the equilibrium concentration which can beprovided due to the danger of adverse effects at the “peak” point. Thepresence of a lag phase may furthermore require supplementary dosingwith repeat injections during the start-up period of depot treatment inorder to maintain a therapeutic dose while the concentrations of activeprovided from the depot are sub-functional. For certain polypeptides inparticular, it would be advantageous to minimise the immediate “burst”effect upon administration of a composition in order to avoid sideeffects such as hypoglycaemia.

One class of peptide hormones which benefits particularly from a very“low burst”, stable in vivo concentration are Somatostatin analogues. Invivo testing suggests that these peptides are particularly beneficialwhen maintained at a steady plasma concentration. This not only suggeststhat a depot composition would be an advantage to avoid “spikes” inconcentration upon administration and/or repeated daily dosing, butfurthermore that such a depot composition should have as flat a releaseprofile as possible during the therapeutic period.

Controlled-release formulations are typically generated frombio-compatible polymers in the form of, for example, implants orinjectable beads. Polymer microsphere formulations must generally beadministered by means of a sizable needle, typically of 20-gauge orwider. This is necessary as a result of the nature of the polymericdosing systems used, which are typically polymer suspensions. It wouldbe an advantage to provide a system of low viscosity, such as ahomogeneous solution, dispersion of fine particles, or L₂ phase, whichcould be administered easily through a narrow needle, thus decreasingthe discomfort of the patient during the procedure. In the case ofdiabetic patients, whether for daytime or nightly use, this ease ofadministration is particularly significant because most patients will befrequently self-administering. Providing a sustained formulation whichcan prevent or reduce the risk of hypoglycemia (especially nocturnalhypoglycemia), but which is sufficiently complex to administer that itrequires treatment by a healthcare professional is unlikely to besuccessful, because the lifestyle disruption involved with such complexadministrations, as well as the costs involved would be too great.Providing a formulation which can be self-administered, and which issufficiently straightforward and painless to administer that patientcompliance is not adversely affected is greatly needed for suchsituations.

The manufacture of PLGA microbeads and suspensions is additionally aconsiderable difficulty with certain existing depot systems. Inparticular, since the beads are particulate, and polymers clogmembranes, they cannot generally be sterile-filtered and furthermore,since the PLGA copolymer melts at around 40° C., they cannot beheat-treated for sterility. As a result, a complex manufacturing processmust all be conducted under conditions of high sterility.

Further issues with biodegradable polymer microspheres include complexreconstitution prior to injection and limited storage stability, dueboth to aggregation and degradation of the delivery system and/oractive.

A lipid-based, slow-release composition is described in WO2006/131730for GLP-1 and analogues thereof. This is a highly effective formulation,but the concentration of active agent which can be included in theformulation is limited by its solubility. Evidently, a higherconcentration of active agent allows for the possibility of longerduration depot products, products maintaining a higher systemicconcentration, and products having a smaller injection volume, all ofwhich factors are of considerable advantage under appropriatecircumstances. It would thus be of considerable value to establish a wayby which higher concentrations of active agents could be included in alipid-based depot formulation.

The present inventors have now established that by providing apre-formulation comprising at least one neutral diacyl glycerol and/or atocopherol, at least one phosphatidyl choline, at least onebiocompatible organic mono-alcoholic solvent, at least one polarsolvent, at least one peptide active agent and optionally at least oneantioxidant in a low viscosity phase, such as molecular solution or L₂(reversed micellar) phase, a pre-formulation may be generated addressingmany of the shortfalls of known depot formulations, and which may beapplied to provide a controlled release of peptide active agent. By useof specific components in carefully selected ratios, and in particularwith a mixture of an alcohol and a polar solvent, a depot formulationcan be generated having a combination of properties exceeding theperformance of even the known lipid controlled-release compositions.

In particular, the pre-formulation shows a highly advantageous releaseprofile, is easy to manufacture, may be sterile-filtered, has lowviscosity (allowing easy and less painful administration typicallythrough a narrow needle), allows a high level of bioactive agent to beincorporated (thus potentially allowing a smaller amount of compositionand/or active agent to be used), requires shallow injection and/or formsa desired non-lamellar depot composition in vivo having a “non-burst”release profile. The compositions are also formed from materials thatare non-toxic, biotolerable and biodegradable, which can be administeredby i.m., or s.c. and are suitable for self-administration. Thepre-formulation may additionally have a very low level of irritation oninjection and in preferred cases causes no irritation at the injectionsite (including transient irritation).

Certain of the formulations of the present invention generate anon-lamellar liquid crystalline phase following administration. The useof non-lamellar phase structures (such as liquid crystalline phases) inthe delivery of bioactive agents is now relatively well established. Amost effective lipid depot system is described in WO2005/117830, and ahighly preferred lipid depot is described in that document. However,there remains scope for achieving depot formulations having improvedperformance in several respects.

Advantages of the compositions of the present invention over polymerformulations, such as PLGA spheres, include the ease of manufacture(including sterilization), handling and use properties combined with lowinitial release (“non-burst profile”) of active agent. This may bedefined such that the area under a plasma concentration against time thecurve during the first 24 hours of a one-month dosing period is lessthan 20% of the area under the curve for the entire curve (measured orextrapolated from time 0 to infinity or from time 0 to the last samplingtime point), more preferably less than 15% and most preferable less than10%. This applies particularly to the acyl saccharide and lipid aspectsof the invention and is discussed in more detail in WO 2005/117830.Furthermore, it may be defined such that the maximum plasmaconcentration of active agent in vivo following injection of thepre-formulation (Cmax) is no more than 10 times, preferably no more than8 times and most preferably no more than 5 times the average plasmaconcentration during the therapeutic period (Cave).

SUMMARY OF THE INVENTION

The present invention provides a pharmaceutical formulation comprisingan appropriate combination of lipid excipients, organic alcoholicsolvent, polar solvent, peptide active agent and certain optionalcomponents, that can be used as a depot-precursor formulation (referredto herein for brevity as a pre-formulation) to address one or more ofthe needs described above.

In a first aspect, the invention therefore provides a pre-formulationcomprising a low viscosity mixture of:

a. 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b.20-80 wt. % of at least one phosphatidyl choline (PC); c. 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; d. up to 20wt. % polar solvent e. at least one peptide active agent; f. optionallyat least one antioxidant;wherein the ratio of components a:b is in the range 40:60 to 54:46;wherein the pre-formulation forms, or is capable of forming, at leastone liquid crystalline phase structure upon contact with excess aqueousfluid.

Such compositions will preferably comprise GDO, ethanol, water/propyleneglycol and/or EDTA as components a), c), d) and f) respectively.Component e) is preferably at least one somatostatin analogue, asdescribed herein.

In a second embodiment, the invention correspondingly provides a processfor the formation of a pre-formulation suitable for the administrationof a peptide bioactive agent to a (preferably mammalian) subject, saidprocess comprising forming a low viscosity mixture of:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; d) up to 20wt. % polar solvent e) at least one peptide active agent; f) optionallyat least one antioxidant;wherein the ratio of components a:b is in the range 40:60 to 54:46;and dissolving or dispersing at least one peptide active agent(preferably a somatostatin analogue) in the low viscosity mixture, or inat least one of components a), b), c), d) and optionally f) prior toforming the low viscosity mixture. Such a pre-formulation will typicallybe one as described herein.

The preformulations are highly useful for the controlled and sustainedrelease of peptide active, especially those requiring or benefiting froma very flat release profile and/or minimal “burst” upon administration.In a corresponding embodiment, the invention therefore provides for theuse of a low viscosity mixture of:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; d) up to 20wt. % polar solvent e) at least one peptide active agent; f) optionallyat least one antioxidant;wherein the ratio of components a:b is in the range 40:60 to 54:46;in the manufacture of a pre-formulation for use in the sustainedadministration of said peptide active agent. Such a low viscositymixture will preferably be one described herein.

The peptide active agents in the formulations of the present inventionare preferably pharmaceutically active. That is to say that they providea therapeutic, palliative and/or prophylactic effect when administeredto a suitable subject (typically being one in need of such an effect).IN a further embodiment, the invention therefore provides a method forthe treatment of a human or non-human mammalian subject comprisingadministering to said subject a pre-formulation as described herein.

Such a method may be for the treatment of a human or non-human mammaliansubject in need thereof to combat, (e.g. cure, improve, prevent orameliorate the symptoms of) at least one condition selected fromacromegaly, cancers, carcinomas, melanomas, tumours expressing at leastone somatostatin receptor, sst(2)-positive tumours, sst(5)-positivetumours, prostate cancers, gastro-entero-pancreatic neuroendocrine (GEPNE) tumours, carcinoid tumours, insulinomas, gastrinomas, vasoactiveintestinal peptide (VIP) tumours and glucagonomas, elevated growthhormone (GH), elevated insulin-like growth factor I (IGF-I), varicialbleeding (especially espohageal), chemotherapy induced gastro intestinalproblems (such as diarrhea), lymphorrhea, diabetic retinopathy, thyroideye disease, obesity, pancreatitis, and related conditions. Such methodsare particularly applicable where component e) is at least onesomatostatin analogue, as described herein. The preformulations asdescribed herein for use in such methods form a further aspect of theinvention.

Correspondingly, in a further aspect, the present invention provides theuse of a low viscosity mixture of:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; d) up to 20wt. % polar solvent e) at least one peptide active agent; f) optionallyat least one antioxidant;wherein the ratio of components a:b is in the range 40:60 to 54:46;in the manufacture of a low viscosity pre-formulation medicament for usein the in vivo formation of a depot for treatment of at least onecondition selected from acromegaly, cancers, carcinomas, melanomas,tumours expressing at least one somatostatin receptor, sst(2)-positivetumours, sst(5)-positive tumours, prostate cancers,gastro-entero-pancreatic neuroendocrine (GEP NE) tumours, carcinoidtumours, insulinomas, gastrinomas, vasoactive intestinal peptide (VIP)tumours and glucagonomas, elevated growth hormone (GH), elevatedinsulin-like growth factor I (IGF-I), varicial bleeding (especiallyespohageal), chemotherapy induced gastro intestinal problems (such asdiarrhea), lymphorrhea, diabetic retinopathy, thyroid eye disease,obesity, pancreatitis, and related conditions. Such uses areparticularly applicable where component e) is at least one somatostatinanalogue, as described herein.

Certain peptide active agents have benefits which are cosmetic ratherthan (or in addition to) therapeutic in nature. Such effects includeweight-loss and/or hunger suppression as well as control over skin orhair pigmentation, hair growth etc. The present invention thereforeadditionally provides a method of cosmetic treatment of a human ornon-human mammalian subject comprising administering to said subject apre-formulation as described herein. Such a cosmetic method willgenerally not be a method of therapy (i.e. will not have therapeutic ormedical benefit).

One of the advantages of the formulations of the present invention overmany other controlled-release compositions is that they are stable tostorage in their final form and thus little or no preparation isrequired at the time of administration. This allows the pre-formulationsto be ready-to-administer and also to be supplied in convenient,ready-to-administer form. In a further aspect, the invention thereforeprovides a pre-filled administration device containing a pre-formulationas described herein. Such a device will generally provide either asingle administration or multiple administrations of a composition whichwill deliver, for example, a dosage of active agent in the range of 1 μgto 5 mg/day.

In a further aspect the invention provides a kit comprising saidadministration device according to the invention.

The kit can optionally contain instructions for subcutaneous orintramuscular administration of said composition. All compositionsdescribed herein are suitable for use in such a kit and may thus becontained therein.

The kits of the invention can optionally include additionaladministration components such as needles, swabs, and the like and willoptionally contain instructions for administration.

BRIEF SUMMARY OF THE ATTACHED FIGURES

FIG. 1a . IVR profile of formulations 911 to 918

FIG. 1b . IVR profile of formulations 1006, 1007, and 1010.

FIG. 2: Peptide Content and Purity (expressed as % of the correspondingvalues obtained for the reference samples stored at <−15° C.) afterstorage of formulations G and H for 7 days at 70° C.

FIG. 3. PK-11-413, dose normalized

FIG. 4: PK-11-425, leuprolide plasma concentration versus time over 21days for formulations 49 and 50.

DETAILED DESCRIPTION OF THE INVENTION

The formulations of the present invention generate a non-lamellar liquidcrystalline phase following administration. The use of non-lamellarphase structures (such as liquid crystalline phases) in the delivery ofbioactive agents is now relatively well established. A most effectivelipid depot system is described in WO2005/117830, and a suitable lipidmatrix for use in the present invention is described in that document,the full disclosure of which is hereby incorporated herein by reference.For a description of the most favourable phase structures of suchformulations, attention is drawn to the discussion in WO2005/117830 andparticularly to page 29 thereof.

All % are specified by weight herein throughout, unless otherwiseindicated. Furthermore, the % by weight indicated is the % of the totalpre-formulation including all of the components indicated herein. Thepre-formulations can optionally consist of essentially only thecomponents indicated herein (including where appropriate additionaloptional components indicated herein below and in the attached claims)and in one aspect consist entirely of such components.

The lipid-based systems described herein comprise lipid components a)and b), plus organic mono-alcoholic solvent (c), polar solvent (d),peptide active agent (e) and optional antioxidant (f) components.

Preferably the pre-formulation according to the invention has an L₂phase structure. Preferably the pre-formulation forms a non-lamellar(e.g. liquid crystalline) phase following administration.

The present inventors have now surprisingly established that byappropriate choice of types, absolute amounts and ratios of lipidcomponents along with a peptide active agent and at least two solventsincluding an alcohol and at least one polar solvent, the releaseproperties of the depot compositions formed from the pre-formulations ofthe invention can be rendered highly advantageous. In particular, byusing a mixture of an alcohol and a polar solvent (especially at theratios close to 1:1 described herein), the advantages of the alcoholsolvent on the release profile can be maintained while other propertiessuch as the comfort on administration and/or the viscosity of theformulation can be improved. Alternatively or in addition to this, therelease profile of the active agent can be made remarkably level, withthe maximum plasma concentration in vivo being only a small multiple ofthe average or even minimum concentration during the dosing period. Suchadvantages apply even in comparison with other lipid depot compositions,which in themselves offer previously unobtainable standards incontrolled release.

It is important, particularly with certain peptide active agents, suchas somatostatin analogues, to control the peak concentration (Cmax) ofdrug in the plasma to a level equal to or less than that tolerable tothe subject, for example to avoid side-effects such as flushing orsevere nausea, while providing or achieving a therapeutically effectivelevel over the desired period of release. Generally, the averageconcentration during the period of release before the next dose isadministered, Cave, falls within the therapeutic range. Control over themaximal (Cmax) and minimum (Cmin) concentrations is also important inorder to achieve the desired treatment over time. In one embodiment, theinitial burst is not the Cmax of the release profile.

Whether or not the initial burst is also the Cmax, preferably theCmax/Cave ratio is less than 50, preferably less than or equal to 15,more preferably less than or equal to 10, even more preferably less thanor equal to 5. Furthermore, it is preferred that the Cave/Cmin ratio isnot more than 50, preferably less than or equal to 15, more preferablyless than or equal to 10, even more preferably less than or equal to 5.Cmax is defined as is known in the art, as the peak or maximal plasmaconcentration observed during the period of release before the next doseis administered and Cave is defined as the average plasma concentrationduring that period of release. Cmin is correspondingly the minimalconcentration during that period. Cave can be calculated by calculatingthe drug present in the plasma as area under the curve (AUC) over theselected period of time, generally the entire period of release beforethe administration of the next dose, and dividing by that period oftime.

Component a)—Diacyl Glycerol

Preferable ranges for component a) are 20-80 wt. %, preferably 30-70 wt.%, more preferably 33-60% (e.g. 43-60%), particularly 38 to 43%.Preferable ranges of component b) are 20-80 wt. %, preferably 30-70 wt.%, more preferably 33-55% (e.g. 35-55%), particularly 38 to 43%.

Ratios of a:b are typically 40:60 to 70:30, preferably 45:55 to 55:45and more preferably 40:60 to 54:46. Ratios of around 50:50 (e.g. ±2) arehighly effective.

Component “a” as indicated herein is preferably at least one diacylglycerol (DAG) and thus has two non-polar “tail” groups. The twonon-polar groups may have the same or a differing number of carbon atomsand may each independently be saturated or unsaturated. Examples ofnon-polar groups include C₆-C₃₂ alkyl and alkenyl groups, which aretypically present as the esters of long chain carboxylic acids. Theseare often described by reference to the number of carbon atoms and thenumber of unsaturations in the carbon chain. Thus, CX:Z indicates ahydrocarbon chain having X carbon atoms and Z unsaturations. Examplesparticularly include lauroyl (C12:0), myristoyl (C14:0), palmitoyl(C16:0), phytanoyl (C16:0), palmitoleoyl (C16:1), stearoyl (C18:0),oleoyl (C18:1), elaidoyl (C18:1), linoleoyl (C18:2), linolenoyl (C18:3),arachidonoyl (C20:4), behenoyl (C22:0) and lignoceroyl (C24:9) groups.Thus, typical non-polar chains are based on the fatty acids of naturalester lipids, including caproic, caprylic, capric, lauric, myristic,palmitic, phytanic, palmitolic, stearic, oleic, elaidic, linoleic,linolenic, arachidonic, behenic or lignoceric acids, or thecorresponding alcohols. Preferable non-polar chains are palmitic,stearic, oleic and linoleic acids, particularly oleic acid.

Mixtures of any number of diacyl lipids may be used as component a).Preferably this component will include at least a portion of C18 lipids(e.g. DAG having one or more C18:0, C18:1, C18:2 or C18:3 non-polargroups), such as glycerol dioleate (GDO) and/or glycerol dilinoleate(GDL). A highly preferred example is DAG comprising at least 50%,preferably at least 80% and even comprising substantially 100% GDO.

Since GDO and other diacyl glycerols are products derived from naturalsources, there is generally a certain proportion of “contaminant” lipidhaving other chain lengths etc. In one aspect, GDO as used herein isthus used to indicate any commercial grade of GDO with concomitantimpurities (i.e. GDO of commercial purity). These impurities may beseparated and removed by purification but providing the grade isconsistent this is rarely necessary. If necessary, however, “GDO” may beessentially chemically pure GDO, such as at least 80% pure, preferablyat least 85% pure and more preferably at least 90% pure GDO.

Component b)—Phosphatidyl Choline

Component “b” in the preferred lipid matrices of the present inventionis at least one phosphatidyl choline (PC). As with component a), thiscomponent comprises a polar head group and at least one non-polar tailgroup. The difference between components a) and b) lies principally inthe polar group. The non-polar portions may thus suitably be derivedfrom the fatty acids or corresponding alcohols considered above forcomponent a. As with component a), the PC will contain two non-polargroups. Again, C18 groups are preferred and may be combined with anyother suitable non-polar group, particularly C16 groups.

The phosphatidyl choline portion, even more suitably than any diacylglycerol portion, may be derived from a natural source. Suitable sourcesof phospholipids include egg, heart (e.g. bovine), brain, liver (e.g.bovine) and plant sources including soybean. Such sources may provideone or more constituents of component b, which may comprise any mixtureof phospholipids. Any single PC or mixture of PCs from these or othersources may be used, but mixtures comprising soy PC or egg PC are highlysuitable. The PC component preferably contains at least 50% soy PC oregg PC, more preferably at least 75% soy PC or egg PC and mostpreferably essentially pure soy PC or egg PC.

In one embodiment applicable to all aspects of the invention, componentb) comprises PC. Preferably the PC is derived from soy. Preferably thePC comprises 18:2 fatty acids as the primary fatty acid component with16:0 and/or 18:1 as the secondary fatty acid components. These arepreferably present in the PC at a ratio of between 1.5:1 and 6:1. PChaving approximately 60-65% 18:2, 10 to 20% 16:0, 5-15% 18:1, with thebalance predominantly other 16 carbon and 18 carbon fatty acids ispreferred and is typical of soy PC.

In an alternative but equally preferred embodiment, the PC component maycomprise synthetic dioleoyl PC. This is believed to provide increasedstability and so will be particularly preferable for compositionsneeding to be stable to long term storage, and/or having a long releaseperiod in vivo. In this embodiment the PC component preferably containsat least 50% synthetic dioleoyl PC, more preferably at least 75%synthetic dioleoyl PC and most preferably essentially pure syntheticdioleoyl PC. Any remaining PC is preferably soy or egg PC as above.

Since the pre-formulations of the invention are to be administered to asubject for the controlled release of a peptide active agent, it isimportant that the components are biocompatible. In this regard, thepreferred lipid matrices for use in the pre-formulations of the presentinvention are highly advantageous since both PC and DAGs are welltolerated and are broken down in vivo into components that are naturallypresent in the mammalian body.

Synthetic or highly purified PCs, such as dioleoyl phosphatidy choline(DOPC) are highly appropriate as all or part of component b). Thesynthetic dioleoyl PC is most preferably1,2-dioleoyl-sn-glycero-3-phosphocholine, and other synthetic PCcomponents include DDPC (1,2-Didecanoyl-sn-glycero-3-phosphocholine);DEPC(1,2-Dierucoyl-sn-glycero-3-phosphocholine);DLOPC(1,2-Dilinoleoyl-sn-glycero-3-phosphocholine);DLPC(1,2-Dilauroyl-sn-glycero-3-phosphocholine);DMPC(1,2-Dimyristoyl-sn-glycero-3-phosphocholine);DOPC(1,2-Dioleoyl-sn-glycero-3-phosphocholine);DPPC(1,2-Dipalmitoyl-sn-glycero-3-phosphocholine);DSPC(1,2-Distearoyl-sn-glycero-3-phosphocholine);MPPC(1-Myristoyl-2-palmitoyl-sn-glycero 3-phosphocholine);MSPC(1-Myristoyl-2-stearoyl-sn-glycero-3-phosphocholine); PMPC(1-Palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine);POPC(1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine); PSPC(1-Palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine); SMPC(1-Stearoyl-2-myristoyl-sn-glycero-3-phosphocholine);SOPC(1-Stearoyl-2-oleoyl-sn-glycero-3-phosphocholine); and SPPC(1-Stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine), or any combinationthereof.

In some circumstances, such as the absence of preserving agents such asEDTA, the use of synthetic or highly purified PCs (e.g. DOPC) mayprovide greater stability for the active agent in the formulations. Thusin one embodiment, component b) may comprise (e.g. may comprise at least75%) synthetic or highly purified (e.g. purity >90%) PCs (e.g. DOPC).This may particularly be in the absence of chelating agents such asEDTA. In an alternative embodiment, component b) may comprise (e.g.comprise at least 75%) naturally derived PCs, such as soy PC or egg PC.This will particularly be where at least one stabilising component (suchas an antioxidant, chelator etc) is included in the precursorformulation.

A particularly favoured combination of components a) and b) are GDO withPC, especially GDO with soy PC and/or DOPC. Appropriate amounts of eachcomponent suitable for the combination are those amounts indicatedherein for the individual components in any combination. This appliesalso to any combinations of components indicated herein, where contextallows.

The ratio of components a:b is in the range 40:60 to 54:46. Preferablythe a:b ratio is in the range 45:55 to 54:46, more preferably 47:53 to53:47. Most preferably the a:b ratio is approximately 50:50.

Component c)—Organic Mono-Alcoholic Solvent

Component c) of the pre-formulations of the invention is an organicmono-alcoholic solvent. Since the pre-formulation is to generate a depotcomposition following administration (e.g. in vivo), typically uponcontact with excess aqueous fluid, it is desirable that this solvent betolerable to the subject and be capable of mixing with the aqueousfluid, and/or diffusing or dissolving out of the pre-formulation intothe aqueous fluid. Solvents having at least moderate water solubilityare thus preferred.

Most preferably component c) comprises or consists of ethanol, propanol,ispropanol, or mixtures thereof. Most preferably component c) comprisesor consists of ethanol.

In a preferred embodiment, the solvent is such that a relatively smalladdition to a mixture comprising a) and b) (i.e. preferably below 15%)gives large viscosity reductions, of one order of magnitude or more. Asdescribed herein, the addition of 10% organic mono-alcohol solvent cangive a reduction of two or more orders of magnitude in viscosity overthe solvent-free composition, or over a depot containing only a polarsolvent such as water, or glycerol.

The amount of component c) in the pre-formulation will have aconsiderable effect upon several features. In particular, the viscosityand the rate (and duration) of release will alter significantly with thesolvent level. The amount of solvent will thus be at least sufficient toprovide a low viscosity mixture but will additionally be determined soas to provide the desired release rate. This may be determined byroutine methods in view of the Examples below. Typically a level of 0.1to 35%, particularly 5 to 25% solvent will provide suitable release andviscosity properties. This will preferably be 5 to 16% (e.g. 6 to 14%)and an amount of around 8% (e.g. 8±2%) is highly effective.

As indicated above, the amount of component c) in the pre-formulationsof the invention will be at least sufficient to provide a low viscositymixture (e.g. a molecular solution, see above) of components a), b), c)and d) and optionally f) and will be easily determined for anyparticular combination of components by standard methods.

The phase behaviour may be analysed by techniques such as visualobservation in combination with polarized light microscopy, X-rayscattering and diffraction techniques, nuclear magnetic resonance, andcryo-transmission electron microscopy (cryo-TEM) to look for solutions,L₂ or L₃ phases, or liquid crystalline phases or as in the case ofcryoTEM, dispersed fragments of such phases. Viscosity may be measureddirectly by standard means. As described above, an appropriate practicalviscosity is that which can effectively be syringed and particularlysterile filtered. This will be assessed easily as indicated herein.Typical organic mono-alcoholic solvents suitable for use in theinvention include at least one solvent selected from ethanol, propanol,isopropanol, and benzyl alcohol, particularly ethanol.

A highly preferred combination for components a), b) and c) is soy PC,GDO and ethanol. As indicated above, appropriate amounts of eachcomponent suitable for the combination are those amounts indicatedherein for the individual components, in any combination.

It is preferable that little or none of component c) contains halogensubstituted hydrocarbons since these tend to have lowerbiocompatibility.

Component c) as used herein may be a single solvent or a mixture ofsuitable solvents but will generally be of low viscosity. This isimportant because one of the key aspects of the present invention isthat it provides pre-formulations that are of low viscosity and aprimary role of a suitable solvent is to reduce this viscosity. Thisreduction will be a combination of the effect of the lower viscosity ofthe solvent and the effect of the molecular interactions between solventand lipid composition. One observation of the present inventors is thatthe oxygen-containing solvents of low viscosity described herein havehighly advantageous and unexpected molecular interactions with the lipidparts of the composition, thereby providing a non-linear reduction inviscosity with the addition of a small volume of solvent.

The viscosity of the “low viscosity” solvent component c) (singlesolvent or mixture) should typically be no more than 18 mPas at 20° C.This is preferably no more than 15 mPas, more preferably no more than 10mPas and most preferably no more than 7 mPas at 20° C.

Component d)—Polar Solvent

Some of the particular benefits of the compositions of the presentinvention come through the unexpected finding that the use of an alcoholsolvent in combination with a polar solvent such as a diol or waterallows a significant improvement in the performance of certainlipid-based controlled-release compositions. In particular, the additionof a diol, such as propylene glycol or water has been observed to reducethe viscosity of a lipid/alcohol/active agent formulation withoutadversely affecting the release profile of the active agent and/orallows the proportion of alcohol to be increased without adverselyaffecting the release profile and/or allows an improvement in therelease profile. By “adversely affecting the release profile” isintended to indicate that the ratio of Cmax/Cave is increased and/or theratio of Cmax/Cmin is increased (for example increased by a factor of atleast 1.2). Similarly an improvement in the release profile indicatesthat the ratio of Cmax/Cave and/or Cmax/Cmin is decreased (e.g.decreased by a factor of at least 1.2.)

Although it has previously been suggested that lipid controlled-releasecompositions should be formulated substantially in the absence of water,in order to avoid the conversion to high-viscosity liquid crystallinephases, it has now furthermore been established that a small andcarefully controlled amount of a polar solvent such as water can provideconsiderable benefits. In particular, the inclusion of this polarsolvent (preferably comprising water) allows further improvements incontrolling the initial release of active agent, allows higher stableloading of some peptide active agents, provides faster depot formationand/or provides further reduced discomfort upon injection. Any one ofthese factors potentially provides a significant improvement in thecontext of therapeutic drug delivery, patient health and/or patientcompliance.

The pre-formulations of the present invention must thus also contain apolar solvent, component d). A suitable amount will typically be greaterthan 1% by weight of the pre-formulation, for example 1-30 wt. %,particularly 1.2-20 wt. %, especially 2-18 wt. %. More preferablycomponent d) is present in the range 5-15 wt. %, especially 6-12 wt. %.Component d) is preferably water, propylene glycol or mixtures thereof.In one preferred aspect, the pre-formulations of the invention containethanol as component c) with water and/or propylene glycol as componentd).

In one embodiment the preformulation comprises at least 1.5% (e.g. atleast 4.5%) water as part of component d) (by weight of the totalcomposition) with the remainder being propylene glycol. At least 5%water with the balance of component d) being PG is preferred. Componentd) may comprise or consist of water.

In an alternative embodiment, component d) may comprise or consist ofpropylene glycol.

Preferably the total level of components c) and d) is not more than 35wt. %, preferably not more than 30 wt. %, preferably 10-30 wt. %, mostpreferably 12-25%.

The ratio of components c) and d) will also have potential advantages inthe compositions of the invention. In particular, by inclusion of somepolar solvent which is miscible with the mono-alcohol component(especially water), the slight sensation that may be caused at theinjection site from the alcohol content can be substantially eliminated.Thus, in one embodiment, the ratio of components c):d) may be in therange 30:70 to 70:30, more preferably 40:60 to 60:40. In one embodiment,the amount of alcohol component c) by weight is no greater than theamount of polar solvent d). Ratios of c):d) ranging from 30:70 to 50:50are thus appropriate in such an embodiment. Approximately equal amountsof components c) and d) are highly appropriate.

A highly preferred combination for the lipid matrix aspect is soy PC,GDO, ethanol, and water/propylene glycol or mixtures thereof. Asindicated above, appropriate amounts of each component suitable for thecombination are those amounts indicated herein for the individualcomponents, in any combination.

Component e)—Peptide Active Agent

The pre-formulations of the present invention contain one or morepeptide active agents. Suitable peptide active agents are disclosed anddiscussed in detail in US WO 2006/075124 and the disclosures of thatdocument are incorporated herein by reference. Suitable peptides for usein the necessary peptides may be naturally occurring or derived fromnatural peptides, or may be chemically modified or wholly syntheticpeptide molecules. Any amino acids may be comprised in the peptidesincluding those described herein, and the peptides may be chemically orenzymatically modified.

Typical peptide actives will be in the range of 500 to 100,000 amu inmolecular weight and can evidently include protein active agents. In oneembodiment, the polypeptides can have at least one cationic charge atneutral and/or physiological pH, and most preferably will require atleast one anionic counter-ion at pH 6.5 or above, preferably at pH 7.5or above. This counter-ion will be physiologically acceptable, and maythus be a halide or the ion of a physiologically acceptable acid.Acetate counter ions and/or chloride ions are particularly preferred andtherefore in one embodiment of the invention, the active agent is apeptide acetate and/or chloride.

Examples of suitable classes of peptides include peptide hormones andsynthetic analogues (such as luteinizing-hormone releasing hormone(LHRH) and analogues (eg, leuprorelin, goserelin, buserelin,tryptorelin, degarelix), incretins and incretin mimetics (such as GLP-1& analogues or glucose-dependent insulinotropic peptide (GIP)),glucagon, insulin and analogues, interferons, vasopressins, calcitonins,etc.), cytokines, antibody fragments (FAbs; scVFs), antimicrobialpeptides (g, corticostatins, defensins, histatins), specific targetingpeptides (e.g., as the examples described in Current Opinion Genetics &Development 10, 71-77 (2006)), venom peptides (e.g., conopeptides), andimmunogenic peptides (e.g., fragments of proteins used for vaccinationpurposes).

In one embodiment, LHRH analogues (also known as GnRH analogues) form apreferred group of active agents for use in the present invention.Preferably such peptides will be structurally related to GnRH I, IIand/or III, and/or one or more of the known analogues, including thoselisted here.

Particularly preferred GnRH analogues are constrained peptides of 6 to12 alpha-amino acids, of which particular examples include thoseindicated above, and particularly leuprolide and goserelin, of thesequences indicated above.

In a further embodiment, GLP-1 and its analogues form a furtherpreferred group of active agents. GLP-1 analogues will be peptides,especially of around 30 amino acids, e.g. 20 to 45, especially 25 to 38.Preferably such peptides will be structurally related to GLP-1 and/orone or more of the known analogues, including those listed here. By“GLP-1 analogue”, as used herein is indicated any GLP-1 receptor agonist(or less preferably antagonist), including naturally occurring forms ofGLP-1, either human or from any other species. These analogues arepreferably peptides, peptide derivatives or peptide mimics. Peptidederived GLP-1 agonists are most preferred, especially GLP-1(7-37),GLP-1(7-36)amide, Liraglutide (Novo Nordisk), AVE-010 (ZP10—ZealandPharma—Sanofi-Aventis), TH0318 (TheraTechnologies), CJC-1131(ConjuChem), LY548806 (Lilly), Exenatide. (Byetta, Amylin-Lilly) andtheir derivatives.

In the peptide actives of the present invention, peptides may containonly amino acids selected from those 20 α-amino acids indicated in thegenetic code, or more preferably may contain their isomers and othernatural and non-natural amino acids, (generally α, β or γ amino acids)and their analogues and derivatives.

Amino acid derivatives are especially useful at the termini of thepeptides, where the terminal amino or carboxylate group may besubstituted by or with any other functional group such as hydroxy,alkoxy, carboxy (on the N-terminal end), ester, amide, thio, amido,amino (on the C-terminal end), alkyl amino, di- or tri-alkyl amino,alkyl (by which is meant, herein throughout C₁-C₂₀ alkyl, preferablyC₁-C₁₈ alkyl e.g. methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-,sec- or t-butyl etc.), aryl (e.g phenyl, benzyl, napthyl etc),heteroaryl, or other functional groups, preferably with at least oneheteroatom and preferably having no more than 20 atoms in total, morepreferably no more than 10 and most preferably not more than 6 atoms(optionally excluding hydrogens).

In one preferred embodiment of the present invention, the peptide activeagent will comprise a somatostatin, or any analogue or derivativethereof.

Somatostatin has two active forms produced by alternative cleavage of asingle preproprotein: one of 14 amino acids, the other of 28 aminoacids. Somatostatin 1-14 is a cyclic peptide hormone having the sequenceAla-Gly-Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys (SEQ ID NO: 1),where the two cystine residues are connected by a disulphide bridge togenerate a type II β-turn at the key binding sequence of Phe-Trp-Lys-Thr(SEQ ID NO: 2). Somatostatin is a natural peptide hormone also known asGrowth Hormone Release Inhibiting Factor and has a role as an antagonistof insulin, glucogen and certain other hormones in the release ofsomatotrophin (Human Growth Hormone). The biological half-life ofnatural Somatostatin is very short (1-3 minutes) and so in itself isdifficult to formulate as a viable therapeutic. However, the lipid depotcompositions of the present invention are highly effective forshort-lived active agents and an increasing number of somatostatinanalogues are becoming available with higher activities and/or longerclearance times in vivo.

Somatostatin analogues, such as octreotide, lanreotide, vapreotide,pasireotide (SOM 230) and related peptides, are used or indicated in thetreatment of a variety of conditions where they are typicallyadministered over an extended period.

Octreotide, for example, is the synthetic octa-peptide with sequenceD-Phe-Cys-Phe-D-Trp-Lys-Thr-Cys-Thr-ol (2-7 disulphide bridge) and istypically administered as the acetate salt. Several clinical studiesalso feature the octreotide pamoate. This derivative retains the keyPhe-(D)Trp-Lys-Thr β-turn but, in contrast to the natural hormone, has aterminal half-life of around 1.7 hours. Octreotide is used in treatmentof conditions including carcinoid tumours and acromegaly, and after aninitial dose is typically given over a sustained period of weeks, ormore commonly many months or years. In addition, somatostatin analoguesare indicated in the treatment of many cancers since a wide variety oftumours are found to express somatostatin receptors. Of particularinterest are those expressing the “sst(2)” and/or “sst(5)” receptor.

The most common “simple” formulation of Octreotide is “Sandostatin”®from Novartis. This is a solution for subcutaneous (s.c) injection and a100 μg dose reaches a peak concentration of 5.2 ng/ml at 0.4 hours postinjection. The duration of action can be up to 12 hours but s.c. dosingis generally carried out every 8 hours. Evidently, s.c. injection 3times daily for periods of months or years is not an ideal dosingregime.

In order to avoid the need for multiple daily injections of octreotide,a further formulation is available; “Sandostatin LAR”®, again fromNovartis. This is a formulation of octreotide in poly lactic co-glycolicacid microspheres which, after resuspension, may be administered byintra muscular (i.m.) injection.

Carcinoid tumours are intestinal tumour arising from specialised cellswith paracrine functions (APUD cells). The primary tumour is commonly inthe appendix, where it is clinically benign. Secondary, metastatic,intestinal carcinoid tumours secrete excessive amounts of vasoactivesubstances, including serotonin, bradykinin, histamine, prostaglandins,and polypeptide hormones. The clinical result is carcinoid syndrome (asyndrome of episodic cutaneous flushing, cyanosis, abdominal cramps, anddiarrhea in a patient with valvular heart disease and, less commonly,asthma and arthropathy). These tumours may grow anywhere in thegastrointestinal tract (and in the lungs) with approximately 90% in theappendix. The remainder occurs in the ileum, stomach, colon or rectum.Currently, treatment of carcinoid syndrome starts with i.v. bolusinjection followed by i.v. infusion. When sufficient effect on symptomshas been established, treatment with a depot formulation of octreotideformulated in ploy lactic-co-glycolic acid (PLGA) microspheres isstarted. However, during the first two weeks or more after injection ofthe depot, daily s.c. injections with octreotide are recommended tocompensate for the slow release from the PLGA spheres.

Acromegaly is a rare chronic and insidious hormonal disorder that occurswhen the pituitary gland produces excess growth hormone (GH). It mostcommonly affects middle-aged adults and may lead to premature death.

Diabetes mellitus, hypertension, and increased risk of cardiovasculardisease are the most serious health consequences of acromegaly. Inaddition, patients with acromegaly are at an increased risk ofdeveloping colon polyps, which can become cancerous. The prevalence ofacromegaly is approximately 60 cases per million population, and theincidence is 3.3 new cases per million per year. The word acromegalycomes from the Greek words for “extremities” (acro) and “great”(megaly), because one of the most common symptoms of this condition isabnormal growth of the hands and feet.

Acromegaly is caused by prolonged overproduction of growth hormone (GH)and excessive production of insulin-like growth factor-I (IGF-I). In 98percent of cases, the overproduction of GH is caused by a pituitaryadenoma. The rate of GH production and the aggressiveness of the tumourvary from patient to patient. Generally, more aggressive tumours areseen in younger patients.

Acromegaly is a severe disease often diagnosed late. Morbidity andmortality rates are high, in particular, because of associatedcardiovascular, cerebrovascular, and respiratory disorders andmalignancies.

Treatment of acromegaly is initiated by a period of s.c. injectionsthree times per day (optimal daily dose=300 μg octreotide). After thelast s.c. dose and providing a suitable effect is observed thentreatment with a depot formulation of octreotide formulated in polylactic-co-glycolic acid (PLGA) microspheres is started. Dose adjustmentsare made after measurement of biomarkers (HG and IGF-1), typically afteraround 3 months.

The existing octreotide slow release formulation relies upon awell-established degrading-polymer type of depot formulation. Typicallysuch formulations are based on a biodegradable polymer such poly (lacticacid) (PLA) and/or poly (lactic-co-glycolic acid) (PLGA) and may be inthe form of a solution in an organic solvent, a pre-polymer mixed withan initiator, encapsulated polymer particles or (as in the case ofoctreotide) polymer microspheres.

In one typical embodiment, the peptide active agent (e.g. somatostatinanalogue) will generally be formulated as 0.02 to 12% by weight of thetotal formulation. Typical values will be 0.1 to 10%, preferably 0.2 to8%, more preferably 0.5 to 6% (e.g. 1 to 3%). These levels may beapplied to all aspects of the invention, where context allows. Foroctreotide, a further preferred range is between 0.5 to 4 wt. %, morepreferably 1-3 wt. %, and most preferably 1.5-2.5 wt. %.

In a related embodiment, the peptide active agent may be formulated at alevel which cannot easily be achieved in the absence of the polarsolvent component of the mixture. In such an embodiment, the peptideactive agent (e.g. Somatostatin analogue) content is typically at least0.7%, preferably at least 1%, more preferably at least 1.8% or at least2% by weight of formulation. Levels of at least 3% and at least 4% areachievable with the present invention, as are loading levels up to 8, 10or 12%. Such compositions of the present invention typically not onlycontain a very high level of peptide active agent (especiallySomatostatin analogue, e.g. octreotide), as indicated, but areadditionally stable to storage without loss or degradation of the activeagent for considerable periods, as indicated herein. Such periods willgenerally be at least a month at 25° C. or at least a month at 5° C.,preferably at least 3 months, and more preferably at least 6 months at5° C. or alternatively at 25° C. These degrees of stability areapplicable to all aspects of the invention, where context allows andrelate to stability both of the active agent and of the phase behaviourof the pre-formulation.

In a related embodiment, in the situation where a peptide active agentis highly soluble in the alcohol component, it may be an advantage tolimit this solubility of this agent. Without being bound by theory, itis thought that excessive solubility in this alcohol component mayresult in the alcohol transporting a significant quantity of activeagent out of the depot composition as it forms in vivo. Therefore, inone embodiment of the present invention, the polar solvent is used tocontrol the solubility of the active agent in the preformulation so asaid control of the release profile.

In one embodiment, the peptide active agent may be a peptide which isnot a somatostatin analogue (as defined herein). For example, thepeptide active agent may be a peptide which does not interact as eitheragonist or antagonist at any of the SST(1) to SST(5) receptors(especially the corresponding human receptors).

In one embodiment, the peptide active agent may be a dual receptormodulator, having a somatostatin analogue directly conjugated to areceptor agonist or antagonist for another receptor. These are referredto herein as “dual receptor agonists. Dual receptor agonists asindicated herein are peptide compounds having at least two distinctdomains wherein one domain serves as an agonist for the somatostatinreceptor and another serves as an agonist or antagonist for anotherbiological receptor. Such dual agonists are distinct from a singlenon-specific agonist in that, although the domains may and preferablywill be covalently bound together, the domain serving as somatostatinreceptor agonist resides on a distinct portion of the peptide sequencefrom the domain serving to affect the other receptor. That is to say,the dual agonist is a compound in which a peptide sequence havingsomatostatin receptor function and substantially no function at thesecond receptor is chemically linked (directly or indirectly) to asequence having function at the second receptor and substantially nosomatostatin receptor agonist function.

In one embodiment, the active agent is not a dual amylin receptor/GLP-1receptor agonist compound.

In a further aspect, the present invention therefore provides a methodfor controlling the solubility of a peptide active agent (such as asomatostatin analogue as described herein) in a low viscosity mixturecomprising:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; e) at leastone peptide active agent; f) optionally at least one antioxidant;by inclusion of a polar solvent component d) to form a depot precursorformulation. Use of a polar solvent in such a method forms a furtheraspect.

The pre-formulations and components of the mixture, as well as theirperformance etc will evidently correspond to those described herein forother aspects.

Similarly, the present invention provides a method for improving therelease profile of a peptide active agent (such as a somatostatinanalogue as described herein) from a depot composition formed byinjection of in a low viscosity mixture comprising:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; e) at leastone peptide active agent; f) optionally at least one antioxidant;by inclusion of a polar solvent component d) in said low-viscositymixture to form a depot precursor formulation. Use of a polar solvent insuch a method forms a further aspect.

The pre-formulations and components of the mixture, as well as theirperformance etc will evidently correspond to those described herein forother aspects.

Corresponding methods and uses provide for the reduction ofinjection-site discomfort, reduction of viscosity of thepre-formulation, and/or reduction in initial “burst” release of a lowviscosity mixture comprising:

a) 20-80 wt. % of at least one diacyl glycerol and/or a tocopherol; b)20-80 wt. % of at least one phosphatidyl choline (PC); c) 5-20 wt. % ofat least one biocompatible, organic mono-alcoholic solvent; e) at leastone peptide active agent; f) optionally at least one antioxidant;by inclusion of a polar solvent component d) in said low-viscositymixture to form a depot precursor formulation. Use of a polar solvent insuch a method forms a further aspect.

All of the above uses and methods for improving the various propertiesof the preformulation and/or the resulting depot composition arepreferably applied without negatively affecting the release profile ofthe peptide active agent.

Where the peptide active agent comprises somatostatin analogue, (e.g.octreotide), suitable doses for inclusion in the formulation, and thusthe volume of formulation used, will depend upon the release rate (ascontrolled, for example by the solvent type and amount used, theantioxidant content and so forth) and release duration, as well as thedesired therapeutic level, the activity of the specific agent, and therate of clearance of the particular active chosen. Typically an amountof around 0.05 to 40 mg per week of depot duration, preferably 0.1 to 20mg per week duration (e.g. 1 to 10 mg per week) for a duration of 1 to24 weeks, preferably 2 to 16 (e.g. 3, 4, 8, 10 or 12) weeks. In analternative embodiment the preformulation may be formulated for dosingweekly (e.g. every 7±1 days). A total dose of 0.05 to 250 mg per dosewould be suitable for providing a therapeutic level for between 7 and168 days. This will preferably be 0.1 to 192 mg, e.g. 0.2 to 160 mg, 0.1to 1.6 mg, 20 to 160 mg etc. Evidently, the stability of the active andlinearity of the release rate will mean that the loading to duration maynot be a linear relationship. A depot administered every 30 days mighthave, for example 0.2 to 20 mg or a 90 day depot have 60 to 120 mg ofactive agent (e.g. Somatostatin analogue, e.g. octreotide). Evidentlyalso, the biological half-life of the specific active will beparticularly important. The half-life of somatostatin, is less than 5minutes, and so for sustained release, a relatively large amount (e.g.towards the higher end of the range) will be needed. For an analoguesuch as octreotide, with a much longer half-life, the amount needed willevidently be lower. Appropriate levels for other actives will beestablished easily by those of skill in the art by reference to theknown therapeutic level, the desired duration of action and the volumewhich is to be injected. A good base calculation would be to multiply atypical daily dose of the active agent by the number of day's durationof the depot. The formulation can then be tested for linearity ofrelease and adjusted as appropriate.

It is a remarkable development of the present formulations that veryshort half-life peptide active agents, comprising, e.g. somatostatin andits analogues can be prepared and administered in a depot precursor ofthe present invention, and will provide controlled release over severaldays or even weeks. This is in spite of the remarkably short biologicalhalf-life of the active agent (e.g. less than 1 hour, preferably lessthan 15 minutes, e.g. less than 5 minutes). Thus, in one embodiment, theactive agent has a half-life of less than 1 hour, e.g. less than 15minutes and the preformulation forms a depot which provides sustainedrelease for at least 7 days, preferably at least 14 days, morepreferably at least 28 days.

Like essentially all organic molecules, lipids and biologically activeagents are thermodynamically unstable to oxidation. As a result, manylipid formulations, including those comprising bioactive agents such asAPIs are susceptible to degradation upon storage, especially byoxidation.

In a highly preferred embodiment, the lipid matrix aspect is soy PC,GDO, ethanol, and water/propylene glycol or mixtures thereof, and thepeptide active agent comprises somatostatin or a somatostatin analogue.As indicated above, appropriate amounts of each component suitable forthe combination are those amounts indicated herein for the individualcomponents, in any combination.

Optional Component f)—Antioxidant

Component f) is an antioxidant. Most preferably it is selected fromascorbic acid, ethylenediaminetetraacetic acid (EDTA) and citric acid.

In all aspects of the invention, component f) is typically present at aweight ratio of antioxidant to peptide active agent of 1:50 to 1:1500,preferably 1:100 to 1:1300, and most preferably 1:150 to 1:1250. Sincetypical antioxidants are of lower molecular weight that the peptideactive agents, the proportion by weight of antioxidant may be relativelysmall. For example, with a small molecular weight pH adjuster (e.g. lessthan 500 amu), 0.0001 to 0.5% of the composition may be antioxidant,preferably 0.0005 to 0.2%, more preferably 0.0008 to 0.1%, e.g. 0.001 to0.015%.

Unfortunately, many common antioxidants are not highly compatible withlipid systems. Indeed, the present inventors have previously establishedthat some antioxidants commonly used in previous systems can causeincreased degradation of active agents in a lipid system. This appliesparticularly to peptide active agents. The present inventors havetherefore analysed a variety of potential antioxidant compounds andclasses for use with lipid based matrix systems and have surprisinglyfound that one particular class of antioxidants is unusually well suitedfor use in these systems.

The antioxidant component is generally included in the range 0.0001 to0.5% by weight of the total pre-formulation. Around 0.0005 to 0.015% ofantioxidant (particularly EDTA) is particularly preferred, especially incombination with the other preferred components and ranges indicatedherein above and below.

Stability data using a number of different antioxidants demonstrate thatEDTA antioxidants are surprisingly more efficient than otherantioxidants in suppressing the oxidative degradation of bioactiveagents. EDTA as antioxidant can also show a synergistic effect incombination with the antioxidants of the present invention, inmaintaining the chemical and physical stability of the peptide activeagent and complete pre-formulation. EDTA has a stabilising effect on theactive agent.

By “stabilising” is indicated an increase in solubility ordispensability of a component (especially an active agent) in the depotdelivery system, or alternatively an increase in the stability of thecomposition, especially with regard to the physical and chemicalstability of the dissolved or dispersed active agent. An increase instability may thus be demonstrated by dissolution, dispersion orsuspension of a greater amount of active agent in the presence of theantioxidant than would be achieved by equilibration, such as byagitation for a prolonged period (e.g. 5 days at 25° C.), in the absenceof antioxidant. Equally, an increase in stability may be demonstrated bythe chemical and/or physical stability of a peptide active agent in alipid formulation for a greater period than would be observed in theabsence of an antioxidant. This would preferably be tested underconditions of typical storage, such as 0-5° C., 25° C. and/or ambienttemperature. This is further described herein below.

Optional Additional Components

In one particularly preferred embodiment of the present invention, thecompositions (preformulations and resulting depots) do not includefragmentation agents, such as polyethyleneoxide or poly(ethylene glycol)(PEG) fragmentation agent, e.g. a PEG grafted lipid and/or surfactant.

For example, the copmositions preferably do not include fragmentationagents such as Polysorbate 80 (P80), or other Polysorbates (e.g.Polysorbate 20), PEGylated phospholipids (PEG-lipids such asDSPE-PEG(2000), DSPE-PEG(5000), DOPE-PEG(2000) and DOPE-PEG(5000)),Solutol HS 15, PEGylated fatty acids (e.g. PEG-oleate), blockco-polymers such as Pluronic® F127 and Pluronic® F68, ethoxylated castoroil derivatives (e.g. Chremophores), PEGylated glyceryl fatty acidesters (such as TMGO-15 from Nikko Chemicals) and PEGylated tocopherols(such as d-alpha tocopheryl poly(ethylene glycol) 1000 succinate knownas Vitamin E TPGS from Eastman.

However, the polypeptide active as a powder (e.g. in the kit of theinvention), as well as active agent dissolved in the lipid formulation,may gain stability (both storage and in vivo stability) by certainstabilising additives. Such additives include sugars (e.g. sucrose,trehalose, lactose etc.), polymers (e.g. polyols such as carboxy methylcellulose), amino acids (such as methionine, glutamate, lysine etc.),lipid-soluble acid components such as HCl, anionic lipids and/or surfaceactive agents (such as dioleoyl phosphatidyl glycerol (DOPG),palmitoyloleoyl phosphatidylglycerol (POPG) and oleic acid (OA)).

Single-dose formats must remain stable and potent in storage prior touse, but are disposable after the single use. In one embodiment, asingle dose format is stable at refrigerated conditions (e.g. 0-5° C.)for at least 12 months. Furthermore such a preformulation may be stableat room temperature (e.g. 25° C.) for at least 12 months. Multi-doseformats must not only remain stable and potent in storage prior to use,but must also remain stable, potent and relatively free of bacteria overthe multiple-dose use regimen administration period after the first usein which a seal has been compromised. For this reason multi-dose formatsoften require a anti-microbial or microbial-static agent, e.g.bacteriostatic agent, preservative.

However, the production of preserved pharmaceutical preparationscontaining protein or peptide actives has often proven difficult, aswhen preservatives are used, these give rise to stability problems.Often the proteins are inactivated and aggregates are formed, which maysometimes lead to reported injection site intolerance or immunogenicityto the active. This can be further aggravated by additional excipientsor formulation components.

In one aspect each of the embodiments herein can optionally contain anantimicrobial or microbial-static agent, which includes bacteriostaticagents and preservative. Such agents include benzalkonium chloride,m-cresol, benzyl alcohol or other phenolic preservatives. Typicalconcentrations as known in the art can be used.

However, surprisingly it has been found that the present formulationswith a peptide active agent do not require an additional preservative,anti-microbial or microbial-static agent, e.g. bacteriostatic orbacteriocide or additional amount of such agent to provide a multi-useformat. The formulations as described herein provide a preservativeeffect with an acceptable peptide stability and formulation stability.They can be used for single-dose as well as for multiple-dose use. Inthis regard, preferred formulations herein for multi-use format cancontain ethanol, propylene glycol, citric acid and/or EDTA as described,preferably in sufficient concentrations to not only provide theirprimary benefit as taught herein but also at sufficient concentration,either alone or in any combination, to provide the preservative effectwhile maintaining stability of the active and the formulation.

Additional components above those mentioned as components a) to f) will,where present at all, preferably be present in an amount of 0 to 5%(e.g. 0.01% to 5%) by weight, preferably no more than 2% by weight andmore preferably no more than 1% by weight.

In one embodiment, components a) and b) (allowing for any impurityinherent in the nature of these components) make up at least 95% of thelipid components of the composition. Preferably at least 99% of thetotal lipid content of the pre-formulation consists of components a) andb). Preferably the lipid component of the pre-formulation consistsessentially of components a) and b).

Administration

The pre-formulations of the present invention are generally formulatedto be administered parenterally. This administration will generally notbe an intra-vascular method but will preferably be subcutaneous (s.c.),intracavitary or intramuscular (i.m.). Typically the administration willbe by injection, which term is used herein to indicate any method inwhich the formulation is passed through the skin, such as by needle,catheter or needle-less (needle-free) injector. It is, however, possibleto take advantage of the high loading and other beneficialcharacteristics of the present formulation in non-parenteralapplications, including topical or systemic application to skin, mucousmembranes, nasal, buccal and/or oral cavities. Preferably, suchnon-parenteral administration is for topical use.

Preferred parenteral administration is by i.m or s.c. injection, mostpreferably by deep s.c. injection. An important feature of thecomposition of the invention is that it can be administered both by i.m.and s.c. and other routes without toxicity or significant local effects.It is also suitable for intracavital administration. The deep s.c.injection has the advantage of being less deep and less painful to thesubject than the (deep) i.m. injection used for some current depots andis technically most suitable in the present case as it combines ease ofinjection with low risk of local side effects. It is a surprisingobservation of the present inventors that the formulations providesustained release of active agent over a predictable time period by bothsubcutaneous and intramuscular injection. This therefore allows the siteof injection to be varied widely and allows the dose to be administeredwithout detailed consideration of the tissue depth at the site ofinjection.

The preferred lipid pre-formulations of the present invention providenon-lamellar liquid crystalline depot compositions upon exposure toaqueous fluids, especially in vivo. As used herein, the term“non-lamellar” is used to indicate a normal or reversed liquidcrystalline phase (such as a cubic or hexagonal phase) or the L3 phaseor any combination thereof. The term liquid crystalline indicates allhexagonal, all cubic liquid crystalline phases and/or all mixturesthereof. Hexagonal as used herein indicates “normal” or “reversed”hexagonal (preferably reversed) and “cubic” indicates any cubic liquidcrystalline phase unless specified otherwise. The skilled reader willhave no difficulty in identifying those compositions having appropriatephase behaviour by reference to the description and Examples providedherein, and to WO2005/117830, but the most favoured compositional areafor phase behaviour is where ratio of components a:b are in the regionof 40:60 to 70:30, preferably 45:55 to 55:45 and more preferably 40:60to 54:46. Ratios of around 50:50 (e.g. ±2) are highly preferred, mostpreferably around 50:50.

It is important to appreciate that the pre-formulations of the presentinvention are of low viscosity. As a result, these pre-formulations mustnot be in any bulk liquid crystalline phase since all liquid crystallinephases have a viscosity significantly higher than could be administeredby syringe or similar injecting dispenser. The pre-formulations of thepresent invention will thus be in a non-liquid crystalline state, suchas a solution, L₂ or L₃ phase, particularly solution or L₂. The L₂ phaseas used herein throughout is preferably a “swollen” L₂ phase containinggreater than 5 wt %, preferably greater than 7%, and most preferablygreater than 9% of organic mono-alcoholic solvent (component c) having aviscosity reducing effect. The preformulations of the invention whichare in L₂ phase form one preferred set of preformulations and these willgenerally contain at least 2% water as polar solvent.

As used herein, the term “low viscosity mixture” is used to indicate amixture which may be readily administered to a subject and in particularreadily administered by means of a standard syringe and needlearrangement. This may be indicated, for example by the ability to bedispensed from a 1 ml disposable syringe through a small gauge needle.Preferably, the low viscosity mixtures can be dispensed through a needleof 19 awg, preferably smaller than 19 gauge, more preferably 23 awg (ormost preferably even 27 gauge) needle by manual pressure. In aparticularly preferred embodiment, the low viscosity mixture should be amixture capable of passing through a standard sterile filtrationmembrane such as a 0.22 μm syringe filter. A typical range of suitableviscosities would be, for example, 0.1 to 5000 mPas, preferably 1 to1000 mPas, more preferably 10 to 750 mPas and most preferably 25 to 500mPas at 20° C.

It has been observed that by the addition of small amounts of lowviscosity organic mono-alcoholic solvent, as indicated herein, a verysignificant change in viscosity can be provided. For example, theaddition of only 5% solvent to a lipid mixture can reduce viscosity100-fold and addition of 10% may reduce the viscosity up to 10,000 fold.In order to achieve this non-linear, synergistic effect in loweringviscosity it is important that a solvent of appropriately low viscosityand suitable polarity be employed. Such solvents include those describedherein infra. Preferred low-viscosity mixtures include molecularsolutions, including dispersions of the peptide active agent in amolecular solution of the other components.

Upon administration, the preferred lipid-based pre-formulations of thepresent invention undergo a phase structure transition from a lowviscosity mixture to a high viscosity (generally tissue adherent) depotcomposition. Generally this will be a transition from a molecularmixture, swollen L₂ and/or L₃ phase to one or more (high viscosity)liquid crystalline phases such as normal or reversed hexagonal or cubicliquid crystalline phases or mixtures thereof. Further phase transitionsmay also take place following administration. Obviously, complete phasetransition is not necessary for the functioning of the invention but atleast a surface layer of the administered mixture will form a liquidcrystalline structure. Generally this transition will be rapid for atleast the surface region of the administered formulation (that part indirect contact with air, body surfaces and/or body fluids). This willmost preferably be over a few seconds or minutes (e.g. from 1 second upto 30 minutes, preferably up to 10 minutes, more preferably 5 minutes ofless). The remainder of the composition may change phase to a liquidcrystalline phase more slowly by diffusion and/or as the surface regiondisperses.

Without being bound by theory, it is believed that upon exposure toexcess aqueous fluid, the pre-formulations of the invention lose some orall of the organic solvent included therein (e.g. by diffusion) and takein aqueous fluid from the bodily environment (e.g. the in vivoenvironment). For lipid pre-formulations, at least a part of theformulation preferably generates a non-lamellar, particularly liquidcrystalline phase structure. In most cases these non-lamellar structuresare highly viscous and are not easily dissolved or dispersed into the invivo environment. The result is a monolithic “depot” generated in vivowith only a limited area of exposure to body fluids. Furthermore,because the non-lamellar structure has large polar, apolar and boundaryregions, the lipid depot is highly effective in solubilising andstabilising active agents such as peptides and protecting these fromdegradation mechanisms. As the depot composition formed from thepre-formulation gradually degrades over a period of days, weeks ormonths, the active agent is gradually released and/or diffuses out fromthe composition. Since the environment within the depot composition isrelatively protected, the pre-formulations of the invention are highlysuitable for active agents with a relatively low biological half-life(see above).

By incorporation of at least 10% of a polar solvent (especially at least5% water) into the pre-formulations, it is believed that the rate ofphase transition to a non-lamellar (e.g. liquid crystalline) phase atthe surface of the injected pre-formulation can be enhanced incomparison with compositions containing organic solvents in thesubstantial absence of water. The performance of the resulting depot isthus improved and further control over the release of active agentachieved.

The depot systems formed by the formulations of the present inventionare highly effective in protecting the active agent from degradation andthus allow an extended release period. The formulations of the inventionthus may provide in vivo depots of peptide active agents which requireadministration only once every 5 to 90 days preferably 5 to 60 days,more preferably 6 to 32. Evidently, a longer stable release period isdesirable for patient comfort and compliance, as well as demanding lesstime from health professionals if the composition is not to beself-administered. Where the composition is to be self-administered,patient compliance may be aided by a weekly (e.g. every 7 days,optionally ±1 day) or monthly (e.g. every 28 or 30 days (optionally ±7days) administration so that the need to administer is not forgotten.

A considerable advantage of the depot precursors of the presentinvention is that they are stable homogeneous phases. That is to say,they may be stored for considerable periods (preferably at least 6months) at room or refrigerator temperature, without phase separation.As well as providing advantageous storage and facile administration,this allows for the dose of peptide active agent (e.g. Somatostatinanalogue, e.g. octreotide) to be selected by reference to the species,age, sex, weight, and/or physical condition of the individual subject,by means of injecting a selected volume.

The present invention thus provides for methods comprising the selectionof a dosing amount specific to an individual, particularly by subjectweight. The means for this dose selection is the choice ofadministration volume.

In one preferred aspect, the present invention provides apre-formulation comprising components a), b), c), d), f) and at leastone peptide active agent (e.g. somatostatin analogue, e.g. octreotide)as indicated herein. The amounts of these components will typically bein the range 30-70% a), 30-60% b), 5-20% c) and 0.1-20% d), with thepeptide active agent (e.g. somatostatin analogue, e.g. octreotide)present at 0.01% to 10%, (such as 36-44% a), 36-44% b), 3-18% c) and5-18% d) (preferably including at least 2% water), with the peptideactive agent (e.g. somatostatin analogue, e.g. octreotide) present at 1%to 3%), wherein the ratio of a:b is in the range 40:60 to 54:46.

Typically, component f) is present at an antioxidant to peptide activeagent molar ratio of 1:50 to 1:1500, preferably 1:100 to 1:1300, andmost preferably 1:150 to 1:1250. Since typical antioxidants are of lowermolecular weight than peptide active agent (e.g. somatostatin analogue,e.g. octreotide), the proportion by weight of antioxidant may berelatively small. For example, with a small molecular weight pH adjuster(e.g. less than 500 amu), 0.001 to 5% of the composition may beantioxidant, preferably 0.002 to 2%, more preferably 0.002 to 0.15%,e.g. 0.002 to 0.015%.

The pre-formulations of the present invention are highly advantageous inthat they are stable to prolonged storage in their final “administrationready” form. As a result, they may readily be supplied foradministration either by health professionals or by patients or theircarers, who need not be fully trained health professionals and may nothave the experience or skills to make up complex preparations. This isparticularly important in long-duration, slow-effecting diseases such asdiabetes.

Devices

In a yet further aspect, the present invention provides a disposableadministration device (which is also to include a device component)pre-loaded with a measured dose of a pre-formulation of the presentinvention. Such a device will typically contain a single dose ready foradministration, and will generally be sterile-packed such that thecomposition is stored within the device until administration. Suitabledevices include cartridges, ampoules and particularly syringes andsyringe barrels, either with integral needles or with standard (e.g.luer) fittings adapted to take a suitable disposable needle.

The pre-filled devices of the invention may also suitably be included inan administration kit, which kit also forms a further aspect of theinvention. In a still further aspect, the invention thus provides a kitfor the administration of at least one peptide active agent, said kitcontaining a measured dose of a formulation of the invention andoptionally an administration device or component thereof. Preferably thedose will be held within the device or component, which will be suitablefor i.m. or preferably s.c. administration. The kits may includeadditional administration components such as needles, swabs, etc. andwill optionally and preferably contain instructions for administration.Such instructions will typically relate to administration by a route asdescribed herein and/or for the treatment of a disease indicated hereinabove.

Kits

The invention provides for a pre-filled administration device asindicated herein and a kit as indicated herein comprising apre-formulation as described herein.

In an alternative aspect of the present invention, the “kit” may containat least two vessels, a first containing a low viscosity mixture ofcomponents a) to d), as described here, and a second containing ameasured dose of at least one peptide active agent as described herein.The antioxidant component f) may be formulated with the active agent, ormore preferably as part of the low viscosity mixture, which will thencomprise components a) to d) and f).

Such a “two component kit” may comprise the peptide active agent as apowder formulation in one vial or pre-filled syringe and components a)to d) (and optionally f)) in a second vial or pre-filled syringe. In thecase of two syringes, before injection, the pre-filled syringes areconnected and the powder comprising active agent is mixed with thematrix formulation by moving the syringe barrels back and forth, forminga solution or suspension which is injected. Alternatively, the liquidlipid formulation is drawn from one vial, or is pre-filled into asyringe, and is injected into a vial containing peptide powder. Thisformulation may subsequently be mixed by hand shaking or other suitablereconstitution method (e.g. vortex mixing etc.). The solvent componentmay be present in either or both vessels (e.g. vials or syringes). Wherethe solvent is at least partially constituted with the active agent,this will generally be in the form of a solution or suspension.

In this aspect, the invention therefore provides a two component kitcomprising

i) a first vessel containing a low viscosity mixture of components a) tod) as described herein;ii) a second vessel containing at least one peptide active agent,iii) an antioxidant component f) optionally in a third vessel,preferably in the second vessel, or most preferably in the first vessel;iv) optionally and preferably at least one of:

-   -   1) at least one syringe (which may be one or both of said first        and second vessels);    -   2) a needle for administration, such as those described herein;    -   3) instructions for generation of a composition of the invention        from the contents of the first and second vessels;    -   4) instructions for administration, whereby to form a depot as        described herein.

Preferred Features and Combinations

In combination with the features and preferred features indicatedherein, the pre-formulations of the invention may have one or more ofthe following preferred features independently or in combination:

All proportions indicated herein may optionally be varied by up to 10%of the amount specified, optionally and preferably by up to 5%;

Component a) comprises, consists essentially of or preferably consistsof GDO;

Component b) comprises, consists essentially of or preferably consistsof soy PC;

Component c) comprises, consists essentially of or preferably consistsof a 1, 2, 3 or 4 carbon alcohol, preferably isopropanol or morepreferably ethanol;

Component d) comprises, consists essentially of or preferably consistsof a polar solvent such as water, propylene glycol, or mixtures thereof;

Component f) comprises, consists essentially of or preferably consistsof ascorbic acid, ethylenediaminetetraacetic acid (EDTA), and/or citricacid;

The pre-formulation contains at least one peptide active agent,preferably a Somatostatin analogue such as Octreotide;

The pre-formulation contains at least one somatostatin analogue (asdescribed herein) such as at least one peptide which has agonisticand/or antagonistic effect at at least one of the SST(1)-SST(5)receptors (e.g. in humans).

The pre-formulation does not contain any somatostatin analogue (asdescribed herein); The pre-formulation has a low viscosity as indicatedherein.

The pre-formulation comprises forms a liquid crystalline phase asindicated herein upon in vivo administration.

The pre-formulation generates a depot following in vivo administration,which depot releases at least one active agent at a therapeutic levelover a period of at least 7 days, preferably at least 21 days, morepreferably at least 28 days.

The pre-formulation has a higher loading of peptide active agent (e.g.Somatostatin analogue, e.g. octreotide) than is stable in the sameformulation in the absence of component e).

The pre-formulation has a higher loading of peptide active agent (e.g.Somatostatin analogue, e.g. octreotide) than is obtainable byequilibration at 25° C. of the same formulation in the absence ofcomponent f).

In combination with the features and preferred features indicatedherein, the method(s) of treatment of the present invention may have oneor more of the following preferred features independently or incombination:

The method comprises the administration of at least one formulation withone or more preferred features as indicated above;

The method comprises the administration of at least one formulation asindicated herein by i.m., s.c. or preferably deep s.c. injection;

The method comprises administration by means of a pre-filledadministration device as indicated herein;

The method comprises administration through a needle no larger than 20gauge, preferably smaller than 20 gauge, and most preferably 23 gauge orsmaller;

The method comprises a single administration every 5 to 90 days,preferably 6 to 32 days (for example 7 days or 28-31 days).

In combination with the features and preferred features indicatedherein, the use(s) of the pre-formulations indicated herein in themanufacture of medicaments may have one or more of the followingpreferred features independently or in combination:

The use comprises the use of at least one formulation with one or morepreferred features as indicated above;

The use comprises the manufacture of a medicament for administration ofat least one formulation as indicated herein by i.m., s.c. or preferablydeep s.c. injection;

The use comprises the manufacture of a medicament for administration bymeans of a pre-filled administration device as indicated herein;

The use comprises the manufacture of a medicament for administrationthrough a needle no larger than 20 gauge, preferably smaller than 20gauge, and most preferably 23 gauge or smaller;

The use comprises the manufacture of a medicament for administrationonce every 5 to 90 days, preferably 5 to 60 days, more preferably 6 to32 days.

In combination with the features and preferred features indicatedherein, the pre-filled devices of the invention may have one or more ofthe following preferred features independently or in combination:

They contain a preferred formulation as indicated herein;

They comprise a needle smaller than 20 gauge, preferably no larger than23 gauge;

They contain a single dose of 1 to 2000 mg of peptide active agent (e.g.Somatostatin analogue, e.g. octreotide), preferably 0.1 to 100 mg andmore preferably 1-50 mg, most preferably 5-35 mg

They contain peptide active agent Somatostatin analogue (e.g. octreotideor exenatide) at around 1 to 100 mg.

They contain a homogeneous mixture of a composition of the invention inready-to-inject form.

They contain a formulation of components a) to c) for combination with apeptide active agent whereby to form a preformulation of the invention.

They contain a peptide active agent for combination with a formulationof components a) to c) and optionally e), whereby to form apreformulation of the invention.

They contain a total volume for administration of no more than 5 ml,preferably no more than 3 ml more preferably no more than 1.5 ml.

In combination with the features and preferred features indicatedherein, the kits of the invention may have one or more of the followingpreferred features independently or in combination:

They contain a preferred formulation as indicated herein;

They contain a pre-filled device as indicated herein;

They contain a needle smaller than 20 gauge, preferably no larger than23 gauge;

They contain a single dose of 1 to 200 mg of peptide active agent (e.g.Somatostatin analogue, e.g. octreotide), preferably 1 to 100 mg and morepreferably 1-50 mg;

They contain peptide active agent. Somatostatin analogue, e.g.octreotide, at around 1 to 100 mg;

They contain a “two compartment kit” comprising at least two vesselscontaining a lipid formulation of the invention and a peptide activeagent (e.g. Somatostatin analogue, e.g. octreotide) powder,respectively.

They contain a total volume for administration of no more than 5 ml,preferably no more than 3 ml more preferably no more than 1.5 ml.

They contain instructions for administration by a route and/or at afrequency as indicated herein;

They contain instructions for administration for use in a method oftreatment as described herein.

The Invention will now be further illustrated by reference to thefollowing non-limiting Examples and the attached Figures.

EXAMPLES

Abbreviations OCT(Cl) Octreotide hydrochloride (PolyPeptide Labs., USA)SOM(Ac) Somatostatin 1-14 acetate (PolyPeptide Labs., USA) LEU(Ac)Leuprolide acetate (PolyPeptide Labs., USA) TTA Triptorelin acetate(Bachem, Switzerland) TTP Triptorelin pamoate (Bachem, Switzerland) SPCSoy phosphatidylcholine (Lipoid, Germany) GDO Glycerol dioleate(Danisco, Denmark) DOPC Dioleoyl phosphatidylcholine (NOF, Japan) EtOHEthanol (99.5 vol %, Ph. Eur., USP) PG Propylene glycol (Ph. Eur., USP)

Example 1: Manufacturing of OCT-Containing Products

TABLE 1 Composition of OCT-containing products. Formu- EDTA- lationIngredient OCT(Cl) SPC GDO EtOH PG water ¹⁾ A (wt %) 2.44 43.78 43.785.00 5.00 — B (wt %) 2.44 42.28 42.28 6.50 6.50 — C (wt %) 2.44 45.5345.53 6.50 — — D (wt %) 2.44 38.78 38.78 10.00 — 10.00 E (wt %) 2.4433.78 33.78 15.00 — 15.00 ¹⁾ The concentration in the solution is 0.10mg EDTA/mL; this solution was prepared by mixing 10 mg EDTANa₂ in water.

Depot precursors with the compositions presented in Table 1 weremanufactured by first preparing a peptide stock, by weighing theingredients as described in Table 2 and mixing on a shaking table(250-300 rpm) to homogeneous solutions.

TABLE 2 Preparation of peptide stock solutions. Formulation IngredientOCT(Cl) EtOH PG EDTA-water A (g) 2.44 5.00 5.00 — B (g) 2.44 6.50 6.50 —C (g) 2.44 6.50 — — D (g) 2.56 — — 10.50 E (g) 0.28 — —  1.73

For manufacturing of A, B and C, the following amounts (Table 3) of SPCand GDO were weighted directly into the recipient containing the OCT(Cl)stock solution.

TABLE 3 Amounts of SPC and GDO added for preparation of CAM2029-BP, -BR,and -BU. Formulation Ingredient SPC GDO A (g) 43.78 43.78 B (g) 42.2842.28 C (g) 45.53 45.53

The mixtures were then placed on a shaking table (250-300 rpm) untilhomogeneous solutions were obtained.

For manufacturing of formulation D, a lipid stock was prepared by mixing(shaking table (250-300 rpm)) 88.58 g SPC, 22.84 g EtOH and 88.58 g GDOto a homogeneous solution. The final formulation was then obtained bycombining 87.56 g lipid stock and 12.44 g peptide stock solution andmixing (shaking table (250-300 rpm)) to homogeneous.

For manufacturing of formulation E, a lipid stock was prepared by mixing(shaking table (250-300 rpm)) 4.09 g SPC, 1.82 g EtOH and 4.09 g GDO toa homogeneous solution. The final formulation was then obtained bycombining 8.26 g lipid stock and 1.74 g peptide stock solution andmixing (shaking table (250-300 rpm)) to homogeneous.

Example 2: In-Vitro Release from OCT-Containing Products

Formulations with the composition presented in Table 4 were manufacturedby first preparing the corresponding OCT(Cl) stock solutions in EtOH,EtOH:PG mixture or respectively water (as described in Example 1 above),followed by mixing with the other components until homogeneous solutionswere obtained.

TABLE 4 Composition of OCT-containing products evaluated in theaccelerated in vitro release (IVR) experiment. Formu- lation IngredientOCT(Cl) SPC GDO EtOH PG Water 911 (wt %) 2.43 43.72 43.79 5.02 5.04 912(wt %) 2.43 42.08 42.13 6.56 6.81 913 (wt %) 2.43 41.26 41.23 7.51 7.58914 (wt %) 2.43 45.41 45.60 6.56 916 (wt %) 2.43 33.70 33.73 15.09 15.06917 (wt %) 2.44 38.70 38.68 10.04 10.14 918 (wt %) 3.64 33.08 33.0715.08 15.13 1006 (wt %) 2.00 41.48 41.48 7.55 7.49 1007 (wt %) 2.0036.49 36.49 12.50 12.53 1010 (wt %) 2.00 38.96 38.95 10.12 9.97

Evaluation of accelerated IVR of OCT from each of the formulationspresented above was carried out by injecting approximately 100 mg (±20%)into a glass vial containing 5 mL of phosphate bufferedsaline:acetonitrile 85:15 (v/v) mixture. The vials were sealed, andincubated at room temperature for up to 48 h. Sampling was carried outat different time points from the initiation of the experiment, byslowly pulling out 0.2 mL of the aqueous phase, which was collecteddirectly into a 0.3 mL HPLC polypropylene vial. The analysis wasperformed by HPLC-UV using an analytical column (ACE-5 C18, 50×3.0 mm)with gradient elution (mobile phase A: 0.1 vol. % trifluoroacetic acid(TFA) in water; mobile phase B: 0.1 vol. % in 95 vol. % methanol, 5 vol.% water) and UV detection at 282 nm.

The results obtained are presented in FIGS. 1 (a and b)

Example 3: Stability of OCT-Containing Products with Vs. Without EDTA

A formulation (batch size 110 g) with the compositionOCT(Cl)/SPC/GDO/EtOH 3.74/43.13/43.13/10.00 (all in wt %) wasmanufactured by first dissolving 4.114 g OCT(Cl) in 11.000 g EtOH,followed by sequential addition of 47.433 g SPC and 47.433 g GDO, andmixing to a homogeneous solution (91).

One sample (G) containing 3.37 wt % OCT(Cl) (approximately 2.98 wt % OCTbase) by mixing 0.9 g formulation 91 with 0.1 mg of a solutioncontaining 0.1 wt % of EDTA in HPLC-grade water.

One sample (H) containing 3.37 wt % OCT(Cl) was prepared by mixing 0.9 gformulation 91 with 0.1 mg HPLC-grade water.

The samples were divided each into two aliquots of about 0.4 g/vial; onealiquot/sample was incubated at 70° C., whereas the other was placed at<−15° C. (reference). All samples were analysed after 7 days ofincubation in the above-mentioned conditions by using a normal-phaseHPLC (analytical column LiChrospher Diol 5 μm, 250×3.2 mm) UV/DAD—basedanalytical method for quantification of OCT and relative determinationof OCT-related substances. The presence of EDTA in the water phaseconsiderably improved the stability of OCT in the lipid matrix, as shownin the results presented in FIG. 2.

Example 4: Injectability of OCT-Containing Products

The injectability is here defined as the flow rate of the evaluatedfluid from a syringe (specified by its volume and design) through aneedle (specified by its needle gauge and length) subjected to aconstant force against atmospheric pressure.

For filling purposes, a thick needle was preferred, e.g. an 18 G needle.When the syringe was filled with the necessary amount of formulation,the thick needle was exchanged for the needle to be examined. Bypressing on the plunger with the new needle in place, the entrapped airwas removed. The excess formulation was wiped off with a paper tissueand the starting weight (grams) of the filled syringe was measured. Thesyringe was then mounted in a vertical position using a metallic standwith holder and with the needle facing down. The ejected fluid wascollected directly in a glass vial.

A 20N weight was placed centered on the plunger and the timer wasstarted when the weight and the plunger come into contact. The time toempty the syringe (seconds) was then monitored. After the syringe hasbeen emptied, its final weight (grams) was measured. At least two repeatmeasurements for each sample and type of needle were performed.

The injectability was calculated by use of the following equation:

${Injectability} = {\frac{\left( {{{Starting}\mspace{14mu}{weight}} - {{Final}\mspace{14mu}{weight}}} \right)}{{Injection}\mspace{14mu}{time}}\left( \frac{mg}{s} \right)}$

The injectability of several OCT-containing products is presented inTable 5.

TABLE 5 Injectability (mg formulation/s) of OCT-containing productsthrough 23 G thin-wall (Terumo Neolus NN-2316R), respective 25 Gthin-wall (Terumo Neolus NN-2516R) 16 mm-long needles. The syringes usedwere BD 1 mL plast Luer-Lock (#309628). Injectability (mg/s)Injectability (mg/s) Formulation through 23 G needle through 25 G needle91 71 25 911 49 17 912 86 38 913 126 49 914 32 12 916 252 109 917 126 49918 308 115

Example 5: In Vivo PK Studies in Rats Animals and Source

Male SPF Sprague-Dawley rats (NTAC:SD) from M&B Taconic Europe A/S(Ejby, Denmark) were used in the studies. At arrival the rats were 8 to9 weeks old, with a bodyweight in the range from 275 to 300 g. Anacclimatization period of at least 5 days was allowed before dosing.

Housing

The rats will be kept in pairs in transparent polycarbonate cages(Macrolon® type III; Scanbur BK A/S, Karlslunde, Denmark) with a floorarea of 810 cm². Aspen wooden chopping (Tapvei Aspen Bedding, Tapvei Oy,Kortteinen, Finland) were used for bedding material. Wood wool for nestbuilding (PM 90 L “Bobyggnadsmaterial”, Tapvei) and a piece of wood(“Gnagpinne medium”, Tapvei) were used as environmental enrichment.Complete pelleted rodent diet (Labfor R70, Kimstad, Sweden) and waterwere available ad libitum.

Dosing

The animals were dosed according to Camurus internal standard operatingprocedure (SOP PK12-3). In brief, dosing was performed by subcutaneousinjections between the scapulae under light isofluran anesthesia, usinga 1-mL Luer-lock syringe and a 25-mm 23 G needle.

Blood Sampling

Blood samples were collected from awake animals by sub-lingual bleeding.Sampling time points was pre-dose, and 1 hour, 6 hours, 1 day, 2 days, 5days, 8 days, 14 days, 21 days, 28 days and 35 days after dosing. Bloodwas collected into EDTA-treated test tubes (Capiject 3T-MQK, TerumoMedical Corporation), placed on ice immediately after collection. Aftercentrifugation (approximately 1500×g, at 5° C. for 10 min) the plasmawas transferred new test tubes and stored below −70° C. until analysis.

Bioanalysis

Analysis of OCT—The plasma samples was analysed with the ELISA kitS-1275 (Bachem/Peninsula Laboratories) “Octreotide—EIA Kit, Host:Rabbit, High Sensitivity”, adapted for analysis of OCT in rat EDTAplasma.

Results

Pharmacokinetic (PK) profiles for formulations 91, 911 and 912 are shownin FIG. 3.

Example 6: Manufacturing of Formulations Containing Leuprolide Acetate(Leuprorelin-LEU(Ac))

Leuprolide compositions according to the invention were prepared withthe compositions as indicated in Table 6. The formulations were preparedby first dissolving the LEU(Ac) in the EtOH, WFI and/or PG components,whereafter the lipid components were added sequentially, starting withSPC and followed by GDO. The final formulations were mixed on a shakingtable at 250-300 rpm until clear and homogenous liquid solutions wereobtained. The formulations were finally subjected to sterile filtration(0.2 μm sterile PVDF filter from Millipore) under 2 bar nitrogenpressure.

TABLE 6 Composition (wt %) of leuprolide acetate (LEU(Ac)) formulations.Formulation# LEU(Ac) SPC DOPC GDO EtOH WFI PG 49 2.70¹⁾ 43.65 — 43.6510.00 — — 50 2.70¹⁾ 37.65 — 37.65 12.00 10.00 — 51 1.62²⁾ 38.19 — 38.1912.00 10.00 — 52 2.70¹⁾ 33.65 — 33.65 15.00 15.00 — 53 2.70¹⁾ 42.15 —42.15 6.50 — 6.50 54 2.70¹⁾ 41.15 — 41.15 7.50 — 7.50 55 2.70¹⁾ 38.65 —38.65 10.00 — 10.00 56 1.62²⁾ 41.69 — 41.69 7.50 — 7.50 57 2.70¹⁾ —41.15 41.15 7.50 — 7.50 ¹⁾Corresponding to 25 mg leuprolide acetate permL when corrected for peptide purity and content and formulationdensity. ²⁾Corresponding to 15 mg leuprolide acetate per mL whencorrected for peptide purity and content and formulation density.

Example 7: Manufacturing of Formulations Containing Triptorelin Acetate(TTA) and Triptorelin Pamoate (TTP)

Triptorelin acetate and pamoate compositions according to the inventionwere prepared with the compositions as indicated in Table 7. Theformulations were prepared by first mixing the TTA or TTP in the EtOHand PG components, whereafter the lipid components were addedsequentially, starting with SPC and followed by GDO. The finalformulations were mixed on a shaking table at 250-300 rpm until clearand homogenous liquid solutions were obtained. The formulations werefinally subjected to sterile filtration (0.2 μm sterile PVDF filter fromMillipore) under 2 bar nitrogen pressure.

TABLE 7 Composition (wt %) of triptorelin acetate (TTA) and triptorelinpamoate (TTP) formulations. Formulation# TTA TTP SPC GDO EtOH PG 583.00¹⁾ — 41.00 41.00 7.50 7.50 59 3.00¹⁾ — 38.50 38.50 10.00 10.00 60 —3.50¹⁾ 40.75 40.75 7.50 7.50 61 — 3.50¹⁾ 38.25 38.25 10.00 10.00¹⁾Corresponding to 25 mg triptorelin free base per mL when corrected forpeptide purity and content and formulation density.

Example 8: Manufacturing of Further Formulations Containing OctreotideChloride (OCT(Cl))

Octreotide compositions according to the invention were prepared withthe compositions as indicated in Table 8. The formulations were preparedby first dissolving the OCT(Cl) in the EtOH, WFI and/or PG components,whereafter the lipid components were added sequentially, starting withSPC and followed by GDO. The final formulations were mixed on a shakingtable at 250-300 rpm until clear and homogenous liquid solutions wereobtained. The formulations were finally subjected to sterile filtration(0.2 μm sterile PVDF filter from Millipore) under 2 bar nitrogenpressure.

TABLE 8 Composition (wt %) of octreotide chloride (OCT(Cl))formulations. Formulation# OCT(Cl) SPC GDO EtOH WFI 81 (91)  3.65¹⁾43.18 43.18 10 — 82 2.44²⁾ 43.78 43.78 10 — 83 (917) 2.44²⁾ 38.78 38.7810 10 84 1.46³⁾ 39.27 39.27 10 10 Corresponding to ¹⁾30 mg, ²⁾20 mg, and³⁾12 mg octreotide free base per mL when corrected for peptide purityand content and formulation density.

Example 9: Manufacturing of Formulations Containing Somatostatin 1-14Acetate (SOM(Ac)) and Somatostatin 1-14 Hydrochloride (SOM(Cl))

Somatostatin (1-14) acetate (SOM(Ac)) and hydrochloride (SOM(Cl))compositions according to the invention were prepared with thecompositions as indicated in Table 9. The hydrochloride salt, SOM(Cl),was prepared from the acetate salt via an ion-exchange chromatographyprocess followed by lyophilisation of the peptide solution byfreeze-drying. Complete counter-ion exchange was confirmed by HPLC. Theformulations were prepared by first mixing the lipid components, SPC andGDO, with the EtOH and PG components, followed by mixing on a shakingtable at 250-300 rpm to form homogenous lipid solutions. To the lipidsolution, the respective SOM(Ac) and SOM(Cl) drug powders were added inthe required amount. The final formulations were mixed by end-over-endrotation at ambient room temperature until clear and homogenous liquidsolutions were obtained. The formulations were finally subjected tosterile filtration (0.2 μm sterile PVDF filter from Millipore) under 2bar nitrogen pressure.

TABLE 9 Composition (wt %) of somatostatin 1-14 acetate (SOM(Ac)) andsomatostatin 1-14 hydrochloride (SOM(Cl)) formulations. Formulation#SOM(Ac) SOM(Cl) SPC GDO EtOH PG 9 — 3.00 43.50 43.50 5 5 11 — 4.00 38.0038.00 10 10 14 3.00 — 38.50 38.50 10 10

Example 10: In Vivo Studies of Leuprolide Formulations in Rats

For general aspects, see Example 5. Dosing of the rats was performed bysubcutaneous injection of formulation 49 and 50, respectively (see Table6).

Blood Samples for Pharmacokinetics

Blood for pharmacokinetics were collected pre-dose, and 1 hour, 4 hours,10 hours, 1 day, 2 days, 3 days, 5 days, 7 days, 14 days and 21 daysafter dosing. The factual time points for sampling were calculated asthe difference between time for sampling and time of dosing. A deviationof ±10% from the nominal time was accepted.

Bioanalysis

Analysis of Leuprolide was performed using the (Des-Gly10, D-LEU6,Pro-NHEt9)-LHRH (Leuprolide) high sensitivity EIA kit (S-1174,Bachem/Peninsula Laboratories) adapted for analysis of LEU in rat EDTAplasma.

Results

Pharmacokinetic (PK) profiles for formulations 49 and 50 are shown inFIG. 4.

1. A method of treating acromegaly, comprising administering to apatient in need thereof a lipid composition comprising 20 mg ofoctreotide or a halide thereof or a physiologically acceptable acidthereof, wherein the lipid composition is administered to the patientonce every 28 days±7 days or 30 days±7 days.
 2. The method of claim 1,wherein the lipid composition is administered to the patient once everymonth.
 3. The method of claim 1, wherein the lipid composition isadministered as a unit dose.
 4. The method of claim 1, wherein theoctreotide or the halide thereof or the physiologically acceptable acidthereof is octreotide chloride.
 5. The method of claim 1, wherein theoctreotide or the halide thereof or the physiologically acceptable acidthereof is the sole active agent in the lipid composition.
 6. The methodof claim 1, wherein the lipid composition further comprises glyceroldioleate, phosphatidylcholine, and ethanol.
 7. The method of claim 6,wherein the lipid composition further comprises propylene glycol.
 8. Themethod of claim 1, wherein the lipid composition consists of a lowviscosity mixture of: a) a lipid component consisting of: at least 95wt. % of: a1) 20-80 wt. % of at least one diacyl glycerol and/or atocopherol; and a2) 20-80 wt. % of at least one phosphatidyl choline(PC); and 0-5 wt. % of at least one impurity associated with componentsa1) and/or a2); b) 5-16 wt. % of ethanol; c) 1-12 wt. % of propyleneglycol (PG); d) the octreotide or the halide thereof or thephysiologically acceptable acid thereof; and e) optionally 0.0005-0.2wt. % at least one antioxidant selected from the group consisting ofascorbic acid, ethylenediaminetetraacetic acid (EDTA), and citric acid;wherein the lipid composition has a viscosity of 10-750 mPas at 20° C.;wherein the ratio of components a1:a2 is in the range 40:60 to 54:46;wherein the ratio of components b:c is in the range 30:70 to 60:40;wherein the lipid composition forms, or is capable of forming, at leastone liquid crystalline phase structure upon contact with excess aqueousfluid; and wherein the lipid composition is for subcutaneous injection.9. The method of claim 8, wherein component a1) comprises glyceroldioleate (GDO).
 10. The method of claim 8, wherein component a1) ispresent at a level of 30-40% by weight.
 11. The method of claim 8,wherein component a2) comprises soy PC.
 12. The method of claim 8,wherein component a2) is present at a level of 30-40% by weight.
 13. Themethod of claim 8, wherein the ratio of components a1:a2 is in the range45:55 to 54:46.
 14. The method of claim 8, wherein component b) ispresent at a level of 6-14 wt. %.
 15. The method of claim 8, whereincomponent c) is present at a level of 6-12 wt. %.
 16. The method ofclaim 8, wherein component d) is octreotide, octreotide chloride, oroctreotide acetate.
 17. The method of claim 8, wherein component d) ispresent at a level of 0.1 to 10 wt. %.
 18. The method of claim 8,wherein the value C_(max)/C_(ave) of component d) is reduced by a factorof at least 1.2 relative to administration of a corresponding lipidcomposition in which component c) is absent.
 19. The method of claim 8,wherein component e) is present.
 20. The method of claim 8, wherein theratio of components c:e is in the range 1:50 to 1:1500.
 21. The methodof claim 8, wherein component a1) consists of glycerol dioleate (GDO),component a2) consists of PC, component c) consists of PG, component d)consists of octreotide chloride, and component e) is present.
 22. Themethod of claim 8, wherein components b) and c) are present inapproximately equal amounts.
 23. The method of claim 22, wherein theratio of components b:c is 50:50.
 24. The method of claim 1, comprisingadministering the lipid composition by a pre-filled administrationdevice.
 25. The method of claim 8, wherein the pre-filled administrationdevice is a syringe, a pre-filled syringe, or a needle-less injector.26. The method of claim 1, administering the lipid composition bysubcutaneous injection.
 27. The method of claim 1, comprisingadministering the lipid composition in a volume of about 1 mL.
 28. Themethod of claim 1, wherein the patient has a maximum blood plasmaconcentration (C_(max))/average blood plasma concentration (C_(ave)) ofoctreotide (at steady state) ratio of less than 50, for each once everymonth administration.
 29. The method of claim 1, wherein the patient hasa maximum blood plasma concentration (C_(max))/average blood plasmaconcentration (C_(ave)) of octreotide (at steady state) ratio of lessthan 15, for each once every month administration.
 30. The method ofclaim 1, wherein the patient has an average blood plasma concentration(C_(ave))/minimum blood plasma concentration (C_(min)) of octreotide (atsteady state) ratio of less than 50, for each once every monthadministration.
 31. The method of claim 1, wherein the patient has anaverage blood plasma concentration (C_(ave))/minimum blood plasmaconcentration (C_(min)) of octreotide (at steady state) ratio of lessthan 15, for each once every month administration.
 32. The method ofclaim 1, wherein the patient has an area under a plasma concentrationagainst time curve during the first 24 hours of the one-month dosing ofless than 20% of the area under the curve for the entire curve (measuredor extrapolated from time 0 to infinity or from time 0 to the lastsampling time point), for each once every month administration.
 33. Themethod of claim 1, wherein the patient has an area under a plasmaconcentration against time curve during the first 24 hours of theone-month dosing of less than 15% of the area under the curve for theentire curve (measured or extrapolated from time 0 to infinity or fromtime 0 to the last sampling time point), for each once every monthadministration.
 34. The method of claim 1, wherein the patient has anarea under a plasma concentration against time curve during the first 24hours of the one-month dosing of less than 10% of the area under thecurve for the entire curve (measured or extrapolated from time 0 toinfinity or from time 0 to the last sampling time point), for each onceevery month administration.
 35. The method of claim 1, wherein thepatient self-administers the lipid composition.